Abstract 3802: Micromilieu-dependent transcriptional memory of tumor vessel endothelial cells in colorectal cancer
Authors: Michael Stürzl, Maximilian Fuchs, Meik Kunz, Nicholas Dickel, Bernt Popp, Steffen Uebe, Arif B. Ekici, Carol I. Geppert, Susanne Merkel, Vera S. Schellerer, Elisabeth Naschberger
Journal: Cancer Research (2022)

Abstract Purpose of the study: Tumor microenvironment (TME)-induced plasticity of fibroblasts and immune cells are known to contribute to cancer pathogenesis. In contrast, only little is known about the existence, stability, functions and prognostic potential of different plastic phenotypes of tumor vessel endothelial cells (TECs) in cancer. Recently, transcriptional memory effects have been observed in different cell lines after cytokine stimulation in vitro. From this we developed the hypothesis that isolated TEC may exhibit TME-dependent transcriptional memories, which may provide novel insights into the intratumoral effects on TEC plasticity in cancer. To investigate this, we isolated ultrapure TECs from colorectal cancer (CRC) with prognostically favorable (Th-1-like) or worse (non-Th1-like) immune-micro-environments and analyzed the prescence of TME-associated transcriptional memory patterns. Methods: TECs and as controls normal colon endothelial cells (NECs), PBMCs and tumor cells (isolated by laser-microdissection) from the same CRC patients (n=75) were isolated by FACS-sorting. Ultra high purity (98%) of TECs and NECs was validated by qPCR and immunocytochemistry. Quality validated complete sample sets from 12 patients (Th-1-like, n=6, non-Th1-like, n=6) were compared using exome sequencing, Clariome S chips and EPICmethylation. Integrative bioinformatics was used to identify differential gene sets and to predict their association with prognosis. Results: Differential transcriptional memory patterns were detected in TECs derived from CRC with different prognostic TMEs. Of note, the aggressiveness of the originating tumors was reflected by the functions of the vascular transcriptional memory genes. The transcriptional memory was imprinted by epigenetic DNA methylation and not due to acquired somatic variants in the genome. These results could be successfully confirmed by alternative methods. Most importantly, integrative bioinformatics analyses retrieved seven vascular memory genes (VMG), which were expressed in the TECs in a TME-dependent manner and predicted the outcome of patients in an independent cohort of CRC patients. Conclusion: Our findings demonstrate for the first time a TME-dependent transcriptional memory of TECs, providing a clinically relevant indicator of intra-tumoral TME-dependent vascular plasticity with perspectives for patient stratification in vessel-directed tumor therapies. Citation Format: Michael Stürzl, Maximilian Fuchs, Meik Kunz, Nicholas Dickel, Bernt Popp, Steffen Uebe, Arif B. Ekici, Carol I. Geppert, Susanne Merkel, Vera S. Schellerer, Elisabeth Naschberger. Micromilieu-dependent transcriptional memory of tumor vessel endothelial cells in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3802.